Abstract
Analyzing cell differentiation during development in a complex organism requires the analysis of expression and chromatin profiles in individual cell types. Our laboratory has developed a simple and generally applicable strategy to purify specific cell types from whole organisms for simultaneous analysis of chromatin and expression. The method, termed INTACT for Isolation of Nuclei TAgged in specific Cell Types, depends on the expression of an affinity-tagged nuclear envelope protein in the cell type of interest. These nuclei can be affinity-purified from the total pool of nuclei and used as a source for RNA and chromatin. The method serves as a simple and scalable alternative to FACS sorting or laser capture microscopy to circumvent the need for expensive equipment and specialized skills. This chapter provides detailed protocols for the cell-type specific purification of nuclei from Caenorhabditis elegans.
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Acknowledgements
This work was supported by HHMI, NIH (U01-HG004274), and the Swiss National Science Foundation (PBSKP3-124362).
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Steiner, F.A., Henikoff, S. (2015). Cell Type-Specific Affinity Purification of Nuclei for Chromatin Profiling in Whole Animals. In: Hancock, R. (eds) The Nucleus. Methods in Molecular Biology, vol 1228. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1680-1_1
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DOI: https://doi.org/10.1007/978-1-4939-1680-1_1
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